Cystic fibrosis (CF) is the most common severe autosomal recessive disease in Caucasian individuals. CF is characterized by an imbalance in the movement of salt and water across the airway surface, such that the overlying mucus layer becomes dehydrated, collapses onto the airway wall, and fails to be cleared from the airways. This impaired mucus clearance leads to chronic infection and airways obstruction, and ultimately causes respiratory failure leading to premature death or lung transplantation. Sodium absorption through the epithelial sodium channel (ENaC), which establishes an osmotic driving force for the absorption of airway surface liquid (ASL), is abnormally high in CF patients for unknown reasons. Furthermore, when the activity of ENaC is inhibited in CF, the ASL becomes hydrated and mucus clearance improves. Our preliminary work suggests that imbalances between proteases, which activate the channel, and endogenous protease inhibitors on the luminal airway surface causes sodium hyperabsorption in CF airways. The goal of this K08 application is to provide the PI with essential skills to develop into a successful academic physician and achieve independent scientific investigator status with a focus on airway physiology as it relates to CF. The PI will focus upon the regulation of airway surface liquid volume by ENaC in normal and CF airway epithelium. The primary scientific aims of this proposal are to (i) define the role of the protease/protease inhibitor balance in the physiological regulation of ENaC and ASL depth, (ii) determine which channel activating proteases and protease inhibitors are involved in the regulation of ENaC in the airway, and (iii) define the mechanism by which proteolytic processing of ENaC is altered in CF. To achieve his goal and complete these aims the PI will combine focused graduate course work with state-of-the-art laboratory methods to delineate the electrophysiology and biochemistry of ENaC in primary human airway epithelial cells and relevant heterologous expression systems. Understanding the nature of proteolytic regulation of ENaC activity in normal and diseased airways will ultimately direct the development of novel therapies to mitigate sodium hyperabsorption and thereby restore normal ASL volume and mucus clearance in CF airways. (End of Abstract)